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Ab59776

Manufactured by Abcam
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Ab59776 is a primary antibody product from Abcam. It is designed for use in immunoassay applications.

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Lab products found in correlation

Cd31 pe, by BD (1 mentions) Cd31 fitc, by BD (1 mentions) Cd45 apc, by BD (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Mouse anti human cd45 fitc, by Miltenyi Biotec (1 mentions) Cd15 fitc, by Miltenyi Biotec (1 mentions) Lamp2, by Abcam (1 mentions) Oct4 c 10, by Santa Cruz Biotechnology (1 mentions) Cd34 pe, by Miltenyi Biotec (1 mentions) Cd14 apc, by Miltenyi Biotec (1 mentions) α sma, by Merck Group (1 mentions) Ripa lysis buffer, by Santa Cruz Biotechnology (1 mentions) Sc 5279, by Santa Cruz Biotechnology (1 mentions) A5228, by Merck Group (1 mentions) P0217, by Agilent Technologies (1 mentions) P0260, by Agilent Technologies (1 mentions) E cadherin, by Cell Signaling Technology (1 mentions) Ab14106, by Abcam (1 mentions) Ab12221, by Abcam (1 mentions) Ab33168, by Abcam (1 mentions)

Close spelling variants of this product

Rabbit anti-SOX2 (ab59776) (2 citations)

5 protocols using ab59776

1

Comprehensive Antibody Panel for Cell Analysis

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The following antibodies were used for flow cytometry, cell sorting and immunostainings respectively: mouse anti-human CD34-APC (Miltenyi and BD biosciences), CD34-PE (Miltenyi), mouse anti-human CD45-FITC (Miltenyi), CD45-APC (BD biosciences), CD45-VB (Miltenyi), mouse anti-human HLA ABC-VB (BD biosciences), CD31-FITC (BD biosciences), CD31-PE (BD biosciences), CD14-APC (Miltenyi), CD15-FITC (Miltenyi), mouse anti-human CD-133-APC (Miltenyi), mouse anti-human CD38-PE (BD biosciences), mouse anti-human CD90-FITC (BD biosciences), HLA-PE (BD biosciences), mouse APC isotype control (BD biosciences), mouse FITC isotype control (BD biosciences), mouse PE isotype control and mouse Vioblue isotype control (BD biosciences), OCT4 (C-10, SantaCruz, sc-5279, 1:100), LAMP-2 (1:50, Abcam), α-tubulin mouse IgG (1:500; Sigma), Anti mouse IgG-Cy3 (1:200; Jackson), Anti rabbit-IgG- Dylight 649 (1:200; Jackson), Alexa fluor 488-, 594- or 647-conjugated anti-rabbit or anti-mouse or anti-goat secondary antibodies (1/500, Jackson Immunoresearch), DAPI (5 mg ml−1) (1:2000; Invitrogen).
For ChIP assay experiment, the following antibodies were used: anti-rabbit-IgGs (sc-2027 Santa Cruz Biotechnology), and rabbit anti-SOX2 (ab59776; Abcam). A total of 5 μg of each antibody was used for ChIP experiments.
Pulecio J., Nivet E., Sancho-Martinez I., Vitaloni M., Guenechea G., Xia Y., Kurian L., Dubova I., Bueren J., Laricchia-Robbio L, & Belmonte J.C. (2014). Conversion of human fibroblasts into monocyte-like progenitor cells. Stem cells (Dayton, Ohio), 32(11), 2923-2938.
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2

Western Blot Analysis of Cell Line Markers

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Whole cell lysate protein was extracted by using RIPA Lysis Buffer (Santa Cruz) and sonicated with Branson Snifier 150. The concentration of extracted protein was measured based on Bradford dye-binding method by using Bio-Rad Protein Assay Reagent and Bio-Rad SmartSpecTM 3000 spectrophotometer. Protein samples were fractionated by size on NuPAGE 4–12% Bis-Tris Gel (Novex) by electrophoresis and then transferred to Nitrocellulose Membrane (Protran, Whatman). The probed primary antibodies were detected by using Horseradish Peroxidise (HRP)-conjugated secondary antibodies and the Enhanced Chemiluminescent (ECL) detection system (GE Health). Primary antibodies: α-SMA (A5228, Sigma-Aldrich, 1:1000); SM22α (ab14106, Abcam, 1:1000); Calponin (EP798Y, Abcam, 1:1000); FSP-1 (ab27957, Abcam, 1:500); SOX2 (ab59776, Abcam, 1:500); OCT4 (sc-5279, Santa Cruz, 1:500); KLF4 (sc-20691, Santa Cruz, 1:500); CD144 (ab33168, Abcam, 1:500); CD31 (252253, ABBIOTEC, 1:500); Claudin 5 (352588, life technologies, 1:500); E-Cadherin (3195 P, Cell Signaling, 1:500); SNAI1 (3879 P, Cell Signaling, 1:1000); N-Cadherin (ab12221, Abcam, 1:300); HES5 (Ab5708, Millipore, 1:400); JAG1 (sc8303, Santa Cruz, 1:300); GAPDH (ab8245, Abcam, 1:2000). Secondary antibodies: anti-rabbit (P0217, DAKO, 1:3000); anti-mouse (P0260, DAKO, 1:3000).
Hong X., Margariti A., Le Bras A., Jacquet L., Kong W., Hu Y, & Xu Q. (2017). Transdifferentiated Human Vascular Smooth Muscle Cells are a New Potential Cell Source for Endothelial Regeneration. Scientific Reports, 7, 5590.
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3

Chromatin Immunoprecipitation and Sequencing

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Genomic DNA was crosslinked and extracted as described above MNase-seq. The purified genomic DNA was suspended in 500 μl of sonication buffer (1XPBS, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS, 1 × EDTA free protease inhibitor cocktail), and sonicated with 6 rounds of 30-s on and 30-s off on ice using XL-2000 Misonix sonicator with power output of 7 Watts. It is critical that the average length of the sheared chromatin is about 250 bp, with length ranging from 150–500 bp. The fragmented DNA was immunoprecipitated with 3~5 μg of specified antibody (anti-Sox2: Abcam ab59776, anti-Pol II: Abcam ab817) as described in the above ChIP-seq for HMs. The ChIP'ed DNA was subjected to massively parallel DNA sequencing on Illumina HiSeq2000 platform (San Diego, CA, USA) using 49 bp single end protocol.
Du Y., Liu Z., Cao X., Chen X., Chen Z., Zhang X., Zhang X, & Jiang C. (2017). Nucleosome eviction along with H3K9ac deposition enhances Sox2 binding during human neuroectodermal commitment. Cell Death and Differentiation, 24(6), 1121-1131.
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4

Culturing and Analyzing Stem Cells

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Fetal bovine serum, trypsin, and Hank’s balanced salt solution were purchased from Invitrogen (Carlsbad, CA, USA). Cell culture plates with ultralow attachment surfaces were purchased from Corning Life Sciences (Tewksbury, MA, USA). Neurobasal medium, FBS, B-27 Supplement, penicillin, and streptomycin were purchased from Life Technologies (Grand Island, NY, USA). TrypLE cell detachment solution was purchased from Thermo Fisher Scientific (Waltham, MA, USA). The HCT-15 cell line was obtained from the American Type Culture Collection. Anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibodies were obtained from EMD Millipore (Billerica, MA, USA). M2 anti-FLAG antibodies were obtained from Sigma-Aldrich (Saint Louis, MO, USA). Antibodies against P53 (SC-6243) and OCT3/4 (SC-8628) were purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA). Antibodies against cyclin D1 (ab134175), NANOG (ab109250), SOX2 (ab59776), and TRRAP (ab73546) were purchased from Abcam (Boston, MA, USA). Antibodies against CD44 (5640), MYC-Tag (2287S), and HA-Tag (3724S) were purchased from Cell Signaling Technology (Danvers, MA, USA).
Kang K.T., Shin M.J., Moon H.J., Choi K.U., Suh D.S, & Kim J.H. (2023). TRRAP Enhances Cancer Stem Cell Characteristics by Regulating NANOG Protein Stability in Colon Cancer Cells. International Journal of Molecular Sciences, 24(7), 6260.
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5

RNA Immunoprecipitation for Studying Protein-RNA Interactions

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RIP was performed as previously described (Ng et al., 2012 (link)). In each immunoprecipitation for the RIP assay, 3 μg of the appropriate antibody was used. For the exogenous RIP assay, an anti-Sox2 (Abcam, ab59776) or anti-Nanog (Abcam, ab80892) antibody was used to bind to the RNA following ectopic expression of linc1614 with Sox2 or Nanog in 293T cells. For the native RIP assay, an anti-Sox2, anti-Nanog, anti-Oct4 (Abcam, ab19857), anti-Ezh2 (Abcam, ab3748), or anti-Suz12 (Abcam, ab12073) antibody was used to pull down the RNA associated with the corresponding proteins. The immunoprecipitated RNA was extracted using RNAiso and analyzed via qPCR. The fold enrichment was calculated relative to the corresponding control IgG sample.
Guo X., Wang Z., Lu C., Hong W., Wang G., Xu Y., Liu Z, & Kang J. (2017). LincRNA-1614 coordinates Sox2/PRC2-mediated repression of developmental genes in pluripotency maintenance. Journal of Molecular Cell Biology, 10(2), 118-129.
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